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Anti pkcε

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Anti-PKCε is a protein kinase C (PKC) epsilon-specific antibody. PKC is a family of serine/threonine-specific protein kinases that play important roles in various cellular processes. The Anti-PKCε antibody can be used to detect and study the PKC epsilon isoform.

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Lab products found in correlation

Lsm 780 confocal microscope, by Zeiss (3 mentions) Anti collagen 2, by Abcam (1 mentions) Anti collagen x, by Abcam (1 mentions) Anti pkcδ, by Abcam (1 mentions) Cultureslide, by BD (1 mentions) Duolink in situ detection reagent orange, by Merck Group (1 mentions) Prolong gold with dapi, by Thermo Fisher Scientific (1 mentions) Duolink in situ pla kit, by Olink (1 mentions) Anti bax, by Cell Signaling Technology (1 mentions) Anti nrf2, by Abcam (1 mentions) Anti bcl 2, by Cell Signaling Technology (1 mentions) Anti cleaved caspase 3, by Cell Signaling Technology (1 mentions) Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions) Anti ho 1, by Cell Signaling Technology (1 mentions) Anti histone h3, by Cell Signaling Technology (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Anti β actin, by Cell Signaling Technology (1 mentions) Hoechst 33342, by Merck Group (1 mentions)

4 protocols using anti pkcε

1

Immunohistochemical Analysis of Chondrocyte Markers

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The expression of collagen II, collagen X and PKC was detected using immunohistochemistry. The slices were dewaxed in xylene, dehydrated in graded ethanol and incubated in 3% H2O2 at 37°C for 10 minutes. Next, they were washed in PBS for 5 minutes thrice, boiled in 0.01 M citric acid buffer for antigen retrieval (95°C, 15‐20 minutes) and blocked in goat serum for 10 minutes at 37°C. Slices were then incubated with primary antibody (anti‐Collagen II (1:200), anti‐Collagen X (1:50), anti‐PKC‐ε (1:50) and anti‐PKC‐δ (1:1000), Abcam, USA) at 4°C overnight and with biotin‐labelled secondary antibody (Bioworld) for 30 minutes at 37°C. Slices were counterstained with haematoxylin and observed under a light microscope.
Ge J., Cheng X., Yan Q., Wu C., Wang Y., Yu H., Yang H., Zhou F, & Zou J. (2020). Calcitonin inhibits intervertebral disc degeneration by regulating protein kinase C. Journal of Cellular and Molecular Medicine, 24(15), 8650-8661.
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2

Visualizing PKCε/Dynein Interaction

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The DuoLink in situ PLA kit (Olink Bioscience) with the DuoLink in situ Detection Reagent Orange (Sigma Aldrich) was used to detect PKCε/Dynein interaction according to the manufacturer’s protocols. HeLa cells were grown on 8-well CultureSlides (Falcon) overnight and treated with or without Blu557 for 1h. Cells were subsequently fixed and permeabilized with PHEM buffer and blocked with 3% BSA/PBS. The slides were directly used for the assay and primary antibody mix solution containing anti-PKCε (AbCam) and anti-DyneinIC (Sigma Aldrich) diluted in 3% BSA/PBS was added to each sample and incubated in a humidity chamber for 1h at room temperature. Mouse IgG and Rabbit IgG (Santa Cruz) were used as negative controls. The assay was subsequently performed following the manufacturer instructions. The slides were mounted with ProLong Gold with DAPI (Invitrogen). Images were acquired using Carl Zeiss LSM 780 confocal microscope equipped with X63 Plan-APOCHROMAT DIC oil-immersion objective and analyzed and serial 1μm Z-sections were taken using ZEN image analysis software. Z-sections were summed and PKCε/Dynein interaction was quantified counting the number of signals each field counted using Image J.
Martini S., Soliman T., Gobbi G., Mirandola P., Carubbi C., Masselli E., Pozzi G., Parker P.J, & Vitale M. (2017). PKCε Controls Mitotic Progression by Regulating Centrosome Migration and Mitotic Spindle Assembly. Molecular cancer research : MCR, 16(1), 3-15.
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3

Immunohistochemical Analysis of PKCε and STAT3

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After transcardial perfusion 7 days after FCA administration with 4% paraformaldehyde, the spinal cords were removed from the rats, postfixed, and dehydrated. Transverse frozen sections (10 μm) prepared from OCT-embedded tissues were incubated overnight with rabbit polyclonal anti-PKCε (Abcam) and mouse monoclonal anti-STAT3 (Cell Signaling Technology). The proteins in the tissues were visualized using a DMIL LED scanning microscope (Leica Microsystems GmbH, Wetzlar, Germany). Primary or secondary antibodies were omitted to ensure staining specificity. The data from three to four sections per rat (n = 6/group) were analyzed.
Li X., Zhou B., Yang H., Yang X., Zhao Z., Pan Z., Liao X., Jian W., Liu Y., Lu H., Xue Q., Luo Y., Yu B., Huang H., Ma D, & Liu Z. (2022). Phosphorylation at Ser 727 Increases STAT3 Interaction with PKCε Regulating Neuron–Glia Crosstalk via IL-6-Mediated Hyperalgesia In Vivo and In Vitro. Mediators of Inflammation, 2022, 2782080.
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4

Antioxidant Mechanisms Regulating Apoptosis

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LyA (purity>98%) was isolated and purified as previously reported [16 (link)]. Dulbecco’s modified Eagle’s medium (DMEM) and fetal bovine serum (FBS) were purchased from Gibco (Gaithersburg, MD, USA). The dihydroethidium (DHE), superoxide dismutase (SOD), glutathione peroxidase (GPx), and malondialdehyde (MDA) assay kits, and the cytoplasmic and nuclear protein extraction kits were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). The anti-p-PKCε, anti-PKCε, anti-Nrf2 were purchased from Abcam (Cambridge, MA, USA). The anti-HO-1, anti-Bcl2, anti-Bax, anti-cleaved caspase3, anti-β-actin, anti-histone H3 were purchased from Cell Signaling Technology (Boston, MA, USA). 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT), dimethylsulfoxide, Hoechst 33342 and all other chemicals were purchased from Sigma-Chemical (St. Louis, MO, USA).
Gao K., Liu M., Ding Y., Yao M., Zhu Y., Zhao J., Cheng L., Bai J., Wang F., Cao J., Li J., Tang H., Jia Y, & Wen A. (2019). A phenolic amide (LyA) isolated from the fruits of Lycium barbarum protects against cerebral ischemia–reperfusion injury via PKCε/Nrf2/HO-1 pathway. Aging (Albany NY), 11(24), 12361-12374.
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